The PSWRCE Protein and PCR MS Core will collaborate on the following projects: Specific Aim i - Proteomics Analyses for Burkholderia, Coccidioides, and Reservoir Vaccines Projects Identify protein biomarkers for use as diagnostic or vaccine candidates (from serum, urine, and lung homogenates, with depletion of abundant mouse/human proteins as necessary) Perform comparative proteomics on pre-fractionated samples (e.g., extracellular proteins or cell wall proteins of two different strains of an organism, or wild type vs. mutant) Build blood meal protein library to use to identify host vertebrate proteins in tick nymphs Specific Aim 2 - Development of Diagnostic Assays Use biomarker IDs of Sp. Aim i to develop bioaffinity MS for pathogen identification Use MS to guide development of targeted ELISA or other fluorescence assays (characterize expressed protein antigens, antibodies, and labeled antibodies by MS throughout development of assay) Compare antibodies vs. small ligands for capture of antigens in bioaffinity MS Specific Aim 3 - Ibis TSOOO PCR-MS Biosensor Applied to Reservoir Vaccines, Burkholderia & Coccidioides PCR MS (on mitochondrial DNA) for ID of vertebrate host on which vector has fed (e.g., tick nymph fed on mule deer) and ID of tick-borne pathogenic organism present PCR MS for rapid ID and strain typing of pathogens, as well as confirmation of the presence of toxin genes, antibiotic resistance genes, selected virulence factor genes Specific Aim 4 - Characterization of Protein Complexes (uncompensated synergistic activity) Structurally characterize large protein-protein complexes (e.g., pili involved in virulence, assemblies of bacteriophage tail fiber proteins useful for biosensors) using a one-of-a-kind modified QTOF MS The antigen discovery coupled to support of diagnostic assay development, PCR-MS, bioaffinity MS, and structural characterization of large protein-protein complexes are unique capabilities of the core.